AN IMPROVED SYNTHESIS OF 9-[2-(DIETHOXYPHOSPHONOMETHOXY)ETHYL]ADENINE AND ITS ANALOGUES WITH OTHER PURINE BASES UTILIZING THE MITSUNOBU REACTION

Abstract

Unprotected adenine, 6-chloropurine, 2.6-diaminopurine. and 2-amino-6-chluropurine have been directly coupled with 2-(diethoxyphosphonomethoxy)ethanol under Mitsunobu reaction conditions to provide acyclic phosphonate nucleotide analogues which are intermediates for antiviral agents such as PMEA.     

结核分枝杆菌对宿主巨噬细胞死亡方式的调控

结核分枝杆菌（Mycobacterium tuberculosis,Mtb）感染后能抑制宿主巨噬细胞（M西）的免疫反应,并在其中生存、复制。研究表明Mtb减毒株感染主要诱导宿主Mφ凋亡,凋亡能抑制胞内Mtb的活力;而Mtb毒力株感染能抑制凋亡的完成,诱导Mφ坏死,最终导致Mtb扩散、感染临近细胞。通过对Mtb感染诱导宿主Mφ不同死亡方式的讨论,进一步认识Mtb的致病机制。     

A Contribution to the Pollen Morphology of Camellia (Theaceae)

The pollen morphology and ultrastructure of 20 species of Camellia (Theaceae) representing the four subgenera were examined. The pollen is tricolporate, spherical to slightly oblate or prolate, with scabrate to rugulate exine sculpturing. The tectum is traversed by perforations that vary in diameter. Pollen wall structure is tectate-columellate, the columellae fused to a footlayer. Endexine is present in all of the taxa examined. The greatest variation was observed in pollen size.     

CXCR4嗜性SHIVKU-1病毒静脉感染中国恒河猴初步研究

目的了解SHIVKU一1静脉途径感染中国恒河猴的感染特点及进展规律。方法两只健康中国恒河猴,静脉感染SHIVKU-1病毒,定期采样检测血浆病毒载量、CD4＋／CD8＋比值、CD4＋T细胞绝对数变化和血清中抗SHIVKU-1特异性IgG抗体水平。多色流式技术分析外周血、腹股沟淋巴结和十二指肠粘膜固有层CD4＋T淋巴细胞记忆细胞亚群变化。结果两只实验猴成功感染SHIVKU-1病毒,一直到感染后3个月均保持稳定水平的病毒载量。外周血CD4＋T淋巴细胞下降明显,CD4＋／CD8＋T细胞比值严重倒置。CD4＋Tcm细胞比例在经历了感染早期的下降后,大幅升高,尤其是外周血和淋巴结。CD4＋Tem则在粘膜固有层中增加明显。结论SHIVKU．1静脉途径成功感染了中国恒河猴,为SHIV／中国恒河猴疾病及评价模型的建立奠定了良好的基础,为今后使用此模型评价抗病毒药物或疫苗提供了条件。     

南美香瓜梨离体培养快速复壮繁殖的研究

南美香瓜梨茎段被移植在ＭＳ基本培养基上,其中添加有ＢＡ６．０（ｍｇ／Ｌ以下单位相同）、ＩＢＡ０．２培养４周后增殖到３．８倍。用其嫩叶切块,在含有２,４—Ｄ０．５,ＢＡ０．２５的ＭＳ培养基上先诱导形成绿色愈伤组织,然后转入含ＢＡ６．０和ＩＢＡ０．２的ＭＳ培养基上,产生丛生不定芽。将２．５～３．０ｃｍ的不定芽切下移入含ＩＢＡ０．５与０．２％活性炭,１／２ＭＳ无机盐的培养基上,２５天后诱导生根,移栽成活率达９０％以上。     

利用强启动功能片段提高麦迪霉素4”，-羟基丙酰化酶基因的表达

利用PCR技术将本室克隆到的强启动功能片段取代麦迪霉素丙酰化酶基因(mpt)的启动子或与mpt基因自身启动子串连,获得含mPt重组质粒pCHFPE3和pCHFPE2。用含有这两个质粒的Streptomyces lividans TK24对螺旋霉素进行微生物转化,结果表明,与含有原启动子的mpt.S.lividans TK24(p.WFPE)相比,丙酰螺旋霉素的组分比例分别提高了89.02％和58.53％。含重组质粒pCHFPE2的螺旋霉素产生菌S.spiramyceticus发酵产物中丙酰螺旋霉素的组分也有较大辐度的提高。说明利用该强启动功能片段可以提高麦迪霉素丙酰化酶基因的表达。     

Influence of sucrose concentration and phosphate source on biomass and metabolite accumulation in adventitious roots of Pseudostellaria heterophylla

In this study, we investigated the induction of Pseudostellaria heterophylla adventitious root and the effects of sucrose concentration and phosphate source on biomass increase and metabolites accumulation. These roots were initially cultured in Murashige and Skoog medium for 4 weeks. IBA 3.0 mg L^?1 proved to be the best auxin for inducing adventitious roots and the frequency of adventitious roots induced from roots (100 %) was higher than that from leaves (78 %) and stems (27 %). The medium with 4 % sucrose resulted in the optimum biomass i.e. 1.04 g/flask DW, and the content of saponin and polysaccharides reached the peak i.e. 0.676 and 24.4 %, respectively. With regards to phosphate source, 1.25 mM phosphate concentration was more favorable for biomass of roots (0.87 g/flask of DW), whereas the optimum saponin (0.74 %) and polysaccharides (22.09 %) were achieved with 2.5 mM phosphate. However, the saponin content at 2.5 mM phosphate did not show significant difference from the saponin content at 0.625 mM (0.69 %) or 3.75 mM phosphate (0.69 %).     

鲑鱼生长激素基因分泌型表达质粒的构建

生长激素(GH)是动物垂体前叶分泌的一种多肽类激素.应用分子重组及PCR等技术,构建了一种鲑鱼生长激素基因分泌型表达质粒pOsGH153,使编码鲑鱼生长激素成熟肽的序列克隆在大肠杆菌分泌型表达载体PIN-Ⅲ-ompA内,直接位于编码大肠杆菌外膜蛋白A信号肽序列的下游,在Lpp-Lac杂合启动子控制下,经IPTG诱导,分子量约23 000的鲑鱼生长激素在大肠杆菌中获得高效表达,该产物具有天然鲑鱼生长激素的免疫活性,直接分泌到细胞周质,而信号肽被自动剪除.     

Depressant effects of prostacyclin in human atrial fibers and cardiomyocytes

The aim of this study was to characterize the electropharmacological effects of prostacyclin (PGI₂) in human atrial fibers and cardiomyocytes. Atrial tissues obtained from the hearts of 28 patients undergoing corrective cardiac surgery were used. Transmembrane action potentials were recorded using a conventional microelectrode technique, and twitch force by a transducer. Effects of PGI₂ (1 nM–10 µM) on action potential characteristics and contraction of atrial fibers were evaluated in normal [K]_o (4 mM) and high [K]_o (27 mM) in the absence and presence of cardiotonic agents. In addition, atrial and ventricular myocytes were isolated enzymatically from atrial tissues and hearts of 4 patients undergoing cardiac transplant. The effects of PGI₂ on Na- and Ca-dependent inward currents (I_Na and I_Ca) of cardiomyocytes were tested. In 9 human atrial fibers showing fast-response action potentials (mean dV/dt_max = 101 ± 15 Vs^–1) in 4 mM [K]_o, PGI₂ did not influence dV/dt_max of phase 0 depolarization even at 1 µM. However, at a concentration as low as 10 nM, PGI₂ depressed spontaneous rhythms or slow-response action potentials in high-K-depolarized fibers. PGI₂ also depressed delayed afterdepolarizations and aftercontractions induced by cardiotonic agents. In isolated cardiomyocytes, PGI₂ reduced I_Ca but not I_Na. The present findings show that, in human atrial fibers and cardiomyocytes, PGI₂ induces greater depressant effects on the slow-response action potential, I_Ca and triggered activity than on the fast-response action potential. It is suggested that PGI₂ may act through a selective reduction of transmembrane Ca influx.     

微生物源脂肽的生物合成和分子调控

微生物源脂肽具有抑制真菌和细菌的生长、抗病毒和抗肿瘤等多种生物活性,在农业生物防治、临床医疗、环境治理等多种领域具有巨大的应用潜力。然而,低产量一直是影响其推广应用的瓶颈。深入了解脂肽合成的关键因素和调控策略对于提高其产量和纯度至关重要。本文概括了3大家族脂肽surfactin、fengycin和iturin的结构、功能及应用前景,介绍了NRPS和NRPS-PKS两种合成系统的结构域和功能,阐释了脂肽生物合成过程中侧链脂肪酸的合成、脂肪酸的活化及与氨基酸的连接、肽链的延伸和环化三个阶段的模块组装和酶催化活动,以及三大家族脂肽合成操纵子开放阅读框的组成;总结了导入或缺失关键基因、定点突变、模块替换、强启动子替换、修饰前体路径等多种遗传操作对脂肽产量的影响,以及群体感应肽信息素、sigma因子等全局调控因子对脂肽合成基因表达的调节。指出利用多组学联用深入探讨脂肽合成的全局分子调控机制和加强结构域蛋白互作和分子动力学研究是提高脂肽产量和纯度以及创造新脂肽的理论基础,提出了利用基因组装和编辑等合成生物学方法及代谢工程技术提高脂肽产量和挖掘新型脂肽靶向性的可能途径,为推进脂肽的生产和应用进程提供科学参考。